全反式维甲酸对人视网膜色素上皮细胞D407增殖及凋亡的影响

Effect of All-trans Retinoic Acid on the proliferation apoptosis in D407 human retinal pigment epithelial cells

目的研究全反式维甲酸(ATRA)对体外培养条件下的人视网膜色素上皮细胞株(retinal pigmentepithelial cells,RPE)D407增殖及调亡的影响,探讨ATRA诱导D407细胞调亡影响的机制。方法将常规传代培养的D407细胞计数后加入浓度为10μg/mL的ATRA完全培养基作为实验组,并与所设空白对照组相比较,继续培养72h后进行活细胞计数并观察药物对D407细胞的抑制作用。同时观察传代细胞培养24h后ATRA对传代细胞贴壁行为的影响。光镜、电镜下观察经ATRA药物作用后D407细胞形态学的改变,应用流式细胞仪检测实验组和对照组的细胞调亡率。结果体外培养的D407细胞ATRA实验组细胞存活数显著少于空白对照组。ATRA能诱导D407细胞发生调亡现象,药物作用72h后发生调亡细胞多为早期调亡细胞。结论 ATRA能有效地抑制体外培养的D407细胞的增殖和黏壁,发生抑制作用的机制可能与诱导细胞调亡有关。

【Objective】 To investigate the effect of All-trans Retinoic Acid（ATRA） on the proliferation and apoptosis in D407 human Retinal pigment epithelium cells induced apoptosis in vitro.【Methods】 D407 cells were cultured conventionally and divided into experimental group and blank group.The secondary cultured D407 cells were counted and then exchanged with the medium contained 10 μg/mL of ATRA as the tested groups,which were compared with the blank control group with normal medium.Then these cells were counted again after another 72 hours to analyze the inhibitory function by ATRA on both groups.The inhibition of the cultured D407 cells attachment by ATRA after 24 hours culture was studied.Light and electronic microscopy was used to observe morphological changes of D407 cells.The apoptotic rate of D407 cells in both testing group and control group cells were analyzed by flow cytometry.【Results】 The cell counts of the ATRA testing groups was prominently decreased than the blank control group.The most of the apoptotic D407 cells were detectable in early stage after 72 hours ATRA exposure.【Conclusion】 The ATRA can inhibit cultured D407 cells proliferation as well as attachment effectively and its inhibition mechanism may be related to cell apoptosis.